Endocytic mechanisms and toxicity of a functionalized fullerene in human cells.

Derivatized fullerenes could be used in biomedical applications and be suitable vectors for drug delivery due to their small size, large surface area and solubility. However, the interactions of derivatized fullerenes with biological systems and cells are not well understood. A water-soluble fullerene-substituted phenylalanine (Bucky amino acid, Baa) poly-lysine derivative with a FITC label (Baa-Lys(FITC)-(Lys)(8)-OH) was characterized by dynamic light scattering, transmission electron microscopy with negative staining, gel electrophoresis, zeta-potential, and UV/vis spectroscopy. Viability assays depicted the cytotoxicity was time, concentration and assay dependent. A decrease in ATP and glutathione at the high concentrations suggests that reactive oxygen species may be involved. Baa-Lys(FITC)-(Lys)(8)-OH was present near the cell membrane at 15 min and entered into the cytoplasm by 30 min but did not localize in the lysosomes. Endocytic inhibitors were used to investigate the uptake mechanism. These results showed that the endocytic pathways could be mediated by caveolae/lipid rafts and cytoskeletal components. A scavenger receptor inhibitor completely blocked the uptake of Baa-Lys(FITC)-(Lys)(8)-OH, suggesting a specific endocytic pathway was strongly involved in Baa-Lys(FITC)-(Lys)(8)-OH cellular uptake.

[1]  M. Prato,et al.  Fullerene derivatives: an attractive tool for biological applications. , 2003, European journal of medicinal chemistry.

[2]  Nancy A Monteiro-Riviere,et al.  Mechanisms of quantum dot nanoparticle cellular uptake. , 2009, Toxicological sciences : an official journal of the Society of Toxicology.

[3]  N. Monteiro-Riviere,et al.  Limitations and relative utility of screening assays to assess engineered nanoparticle toxicity in a human cell line. , 2009, Toxicology and applied pharmacology.

[4]  H. Yamawaki,et al.  Cytotoxicity of water-soluble fullerene in vascular endothelial cells. , 2006, American journal of physiology. Cell physiology.

[5]  Nancy A. Monteiro-Riviere,et al.  Challenges for assessing carbon nanomaterial toxicity to the skin , 2006 .

[6]  B. Röder,et al.  Intracellular Uptake and Phototoxicity of 31,32‐Didehydrophytochlorin‐fullerene Hexaadducts , 2007, Photochemistry and photobiology.

[7]  D. Carroll,et al.  C60-Fullerenes: detection of intracellular photoluminescence and lack of cytotoxic effects , 2006, Journal of nanobiotechnology.

[8]  Eva Oberdörster,et al.  Ecotoxicology of carbon-based engineered nanoparticles: Effects of fullerene (C60) on aquatic organisms , 2006 .

[9]  M. Akashi,et al.  Targeting of Antigen to Dendritic Cells with Poly(γ-Glutamic Acid) Nanoparticles Induces Antigen-Specific Humoral and Cellular Immunity1 , 2007, The Journal of Immunology.

[10]  Yu-Ying He,et al.  Pristine (C60) and hydroxylated [C60(OH)24] fullerene phototoxicity towards HaCaT keratinocytes: type I vs type II mechanisms. , 2008, Chemical Research in Toxicology.

[11]  J. West,et al.  The Differential Cytotoxicity of Water-Soluble Fullerenes , 2004 .

[12]  Na Zhang,et al.  PLGA nanoparticle--peptide conjugate effectively targets intercellular cell-adhesion molecule-1. , 2008, Bioconjugate chemistry.

[13]  P. Bernier,et al.  Effects of fullerenes and single-wall carbon nanotubes on murine and human macrophages , 2006 .

[14]  Serdar Durdagi,et al.  In Silico Drug Screening Approach for the Design of Magic Bullets: A Successful Example with Anti-HIV Fullerene Derivatized Amino Acids , 2009, J. Chem. Inf. Model..

[15]  M. Lenardo,et al.  Photoconversion of Lysotracker Red to a green fluorescent molecule , 2007, Cell Research.

[16]  S. Kannan,et al.  Poly(amidoamine) dendrimer-drug conjugates with disulfide linkages for intracellular drug delivery. , 2009, Biomaterials.

[17]  N. Monteiro-Riviere,et al.  Trace analysis of fullerenes in biological samples by simplified liquid-liquid extraction and high-performance liquid chromatography. , 2006, Journal of chromatography. A.

[18]  A. Barron,et al.  Fullerene-derivatized amino acids: synthesis, characterization, antioxidant properties, and solid-phase peptide synthesis. , 2007, Chemistry.

[19]  S. Kannan,et al.  Anti-inflammatory and anti-oxidant activity of anionic dendrimer-N-acetyl cysteine conjugates in activated microglial cells. , 2009, International journal of pharmaceutics.

[20]  Nancy A Monteiro-Riviere,et al.  Effects of mechanical flexion on the penetration of fullerene amino acid-derivatized peptide nanoparticles through skin. , 2007, Nano letters.

[21]  M. Ponec,et al.  LDL receptors in keratinocytes. , 1992, The Journal of investigative dermatology.

[22]  P. Elias,et al.  Scavenger Receptor Class B Type I Is Expressed in Cultured Keratinocytes and Epidermis , 2002, The Journal of Biological Chemistry.

[23]  Nancy A Monteiro-Riviere,et al.  Fullerene-based amino acid nanoparticle interactions with human epidermal keratinocytes. , 2006, Toxicology in vitro : an international journal published in association with BIBRA.

[24]  Zoran Markovic,et al.  Distinct cytotoxic mechanisms of pristine versus hydroxylated fullerene. , 2006, Toxicological sciences : an official journal of the Society of Toxicology.

[25]  K. Ausman,et al.  C60 in water: nanocrystal formation and microbial response. , 2005, Environmental science & technology.

[26]  Jianzhong Yang,et al.  A new route to fullerene substituted phenylalanine derivatives. , 2004, Chemical communications.

[27]  A. Barron,et al.  Biological Interactions of Functionalized Single-Wall Carbon Nanotubes in Human Epidermal Keratinocytes , 2007, International journal of toxicology.

[28]  A. Barron,et al.  The use of fullerene substituted phenylalanine amino acid as a passport for peptides through cell membranes. , 2007, Organic & biomolecular chemistry.

[29]  Alok Dhawan,et al.  Stable colloidal dispersions of C60 fullerenes in water: evidence for genotoxicity. , 2006, Environmental science & technology.

[30]  K. Tsujii,et al.  Stable colloidal dispersions of fullerenes in polar organic solvents. , 2001, Journal of the American Chemical Society.